Establishing a papillary craniopharyngioma cell line by SV40LT-mediated immortalization

Pituitary. 2021 Apr;24(2):159-169. doi: 10.1007/s11102-020-01093-5. Epub 2020 Oct 12.

Abstract

Background: Craniopharyngioma represents a troublesome tumor of the intracranial sellar region. There are currently no available well-characterized craniopharyngioma cell lines. This lack of reliable, immortal cell lines is a major reason for the slow progress in fundamental research related to craniopharyngioma.

Methods: We describe the development of an immortal papillary craniopharyngioma (PCP) cell line by transfecting primary PCP cells with the pLenti-simian virus 40 large T antigen(SV40LT).

Results: Three clones have been cultured for more than 14 months so far, while non-transfected cells ceased proliferation within three months of isolation. The established immortal PCP cell lines were identified to have BRAFV600E mutations, while no mutations in tumor suppressor genes were found in primary cells or immortal cells. Immortal cells had higher proliferation rates and formed tumors when implanted in the bran of nude mice. BRAF inhibition in immortal PCP cells altered cell morphology, inhibited cell proliferation and promoted apoptosis.

Conclusion: We successfully developed PCP cell lines by SV40LT-mediated immortalization. These cell lines represent a powerful tool for fundamental and therapeutical studies on craniopharyngioma.

Keywords: Immortalization; Papillary craniopharyngioma (PCP); SV40LT.

MeSH terms

  • Animals
  • Antigens, Viral, Tumor / immunology*
  • Cell Line, Tumor
  • Craniopharyngioma / immunology*
  • Cyclin-Dependent Kinase Inhibitor p16 / genetics
  • Fluorescent Antibody Technique
  • Humans
  • Immunohistochemistry
  • Male
  • Mice
  • Mice, Nude
  • Middle Aged
  • Simian virus 40 / immunology*
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / genetics
  • rho GTP-Binding Proteins / genetics

Substances

  • Antigens, Viral, Tumor
  • CDKN2A protein, human
  • Cyclin-Dependent Kinase Inhibitor p16
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • rho GTP-Binding Proteins